The present invention relates to new denitrifying aerobic bacterial compositions and to aerobic methods for biological treatment of aqueous bodies and aqueous systems polluted by nitrogen waste products.
Water systems commonplace in today""s environment including aquarium aquaculture, streams, rivers, ponds, lakes, and ground water, are all subject to contamination by many inorganic and organic compounds, as well as nitrogen waste components, in particular, ammonium, nitrites and nitrates, which at certain concentrations become highly toxic. Municipal wastewater solutions, for examples raw sewage, and industrial effluents, often contain high levels of ammonium as well as phenols, cyanides, sulfates, thiocyanates, thiosulfates, and other toxic waste compounds. Many spent military explosives contain organonitrate waste, and removing such waste in an economical fashion without contamination of the environment remains a challenge. These nitrogen waste streams are one of the principal major ecological and environmental problems facing mankind, and will continue to pose serious problems as the population grows and waste production increases.
Removing nitrogen waste from contaminated aqueous bodies and wastewater streams is accomplished primarily by bacterial degradation. Specifically, bacteria from the families of Nitrosomas, Nitrobactor, and Pseudomonas are used to digest the nitrogen waste. These bacterial types are members of the well-characterized nitrogen cycle. The bacterial families of the classical nitrogen cycle act to transform nitrogen containing waste products to nitrogen gas and other, offensive odiferous and environmentally contaminating gases. Bacterial cultures from the Nitrosomas and Nitrobactor families convert, via aerobic bioxidative processes, the ammonia or organic amine waste to nitrate waste. At very low or with no oxygen present, pseudomonas bacterial cultures are employed to bioreduce the nitrate waste products and transform such waste into nitrogen gas in combination with methane, sulfur dioxide, hydrogen sulfate and carbon dioxide. This bioreduction by Pseudomonas cultures is accomplished anaerobically. The time required to establish the classical nitrogen cycle using the nitrobacter and pseudomonas families is from about twenty eight days to six weeks, and is dependent upon: (a) temperature; (b) bio mass available for culture; (c) the amount and type of biological platform for colonization; (d) concentration of hydronium ion (pH); (e) amount of dissolved oxygen; and, (f) available food (REF). Optimal conditions for the biological redox processes of removing nitrogen pollutants from water bodies, or wastewater streams requires a fine balance of these factors; limiting one or more results in a significantly less efficient biological processing and thus engendering a substantially longer time to process the waste.
The prior art has many examples of teachings that employ bacterial compositions to accomplish, or aid in accomplishing, the biologically mediated purification of wastewater. For example, denitrifying bacterial compositions are used in combination with solid column packings in the teachings of Francis, U.S. Pat. No. 4,043,936. These compositions are believed to belong to the family of Pseudomonas. Hater, et al U.S. Pat. No. 4,810,385 teaches a wastewater purification process involving bacterial compositions comprising, in addition to non ionic surfactants and the lipid degrading enzymes Lipase, three strains of Bacillus subtillis, 3 strains of Pseudomonas aeruginosa, one strain of Pseudomonas stutzeri, one strain of Pseudomonas putida, and one strain of Eschericia hermanii grown on a bran base. Wong, et. al., U.S. Pat. No. 5,284,587 teaches a bacterial composition, that is in combination with enzymes and a gel support is necessary to achieve satisfactory waste treatment. Bacterial species mentioned in Wong et al are Bacillus subtillis, Bacillus licheniformis, Cellulomonas and acinetobacter lwoffi. Similarly, Wong and Lowe, U.S. Pat. No. 4,882,059 teach a process for biological treatment of wastewater comprising bacterial species that aid in the solubilization of the solid debris. The bacterial species used in the teaching of Wong and Lowe are of the following bacterial types: Bacillus amyloliquefaciens and aerobacter aerogenes. These bacterial types are taught to be employed primarily for solubilization and biodegradation of starches, proteins, lipids and cellulose present in the waste product.
Other patents for treating wastewater streams are Luck, U.S. Pat. No. 3,801,494; Erickson et al U.S. Pat. No. 4,159,944; Bhattacharyya, U.S. Pat. No. 4,271,013; Semp et al U.S. Pat. No. 4,566,469; Li et al, U.S. Pat. No. 5,086,036; and Rusherford, et. al., U.S. Pat. No. 5,223,140.
The shortcomings of the prior art are the extreme sensitivity of the method, not only for the time required to establish the nitrogen cycle, but also for the criticality of the various parameters of the processes, such as pH, temperature, food source and bacterial habitat. Also, a major shortcoming of the prior art are the elaborate mechanical and chemical methods and processes required to assist the bacterial compositions in the purification process. Another major shortcoming of the prior art is that there exists no effective method to significantly reduce or eliminate completely the odiferous side-products that are produced. The gases produced by the classical denitrification biological processes, specifically hydrogen sulfide, as well as others are the source of the foul smell that is one of the most noticeable problems associated with waste treatment, and the most toxic to man and the environment.
There exists a need in treatment of aqueous bodies, wastewater streams and systems and other applications for a more efficacious and faster biological process for converting toxic inorganic and organic amino, nitroso, nitrite, nitro and nitrate so that they are non-toxic to human, animal and marine species.
The present invention is the discovery of a novel composition and method for the aerobic oxidation and reduction of organic and inorganic nitrogen waste, and in particular a unique composition of bacteria and of bacteria and enzymes that, in addition to oxidizing ammonia and nitrites, organic amines and organonitrites, will aerobically reduce nitrates to molecular nitrogen. The proper application of the present invention leads to a significantly improved method for biological processing of aqueous bodies, wastewater streams and systems that overcome many of the shortcomings of the prior art, byxe2x80x94in particular: (1) the elimination of toxic or explosive by-products; (2) reduction of time required to establish the nitrogen cycle (reduced from about 28 days to 1-3 days and to 24 hours); (3) reduction of sensitivity to pH; (4) elimination of the need for side-processes for Aactivation@ or for other enzymes or apparatus to facilitate biological degredative process; and (5) the sensitivity to temperature.
The present invention includes use of a mixture of bacteria selected from and limited to bacillus bacteria for the purification of contaminated aqueous bodies, wastewater systems and wastewater streams. it is a totally aerobic process.
In one aspect the present invention includes a method for the purification of contaminated aqueous bodies, wastewater systems and wastewater streams using a composition of certain combinations of the bacterial families: Bacillus subtillis, Bacillus sphaericus, Bacillus megatarium, Bacillus licheniformis, Enterobacter sakazakii, Bacillus coagulans, Bacillus cereus, Bacillus pasteurii, Bacillus cirroflagellosus, Bacillus pumilus, and optionally with a carbon platform and carbon source, and optionally with the enzymes Cellulase, Amylase, Protease, Lipase, and optionally with selected pancreatic preparations, and kidney microbia compositions.
It is the prime objective of the present invention to provide for a composition and method for bacteria-mediated aerobic reduction of nitrate molelies in aqueous systems.
It is a further object of the present invention to employ novel bacterial compositions comprising the following bacterial families: Bacillus subtillis, Bacillus sphaericus, Bacillus megatarium, Bacillus licheniformis, Enterobacter sakazakii, Bacillus coagulans, Bacillus cereus, Bacillus pasteurii, Bacillus cirroflagellosus, Bacillus pumilus,that provide for the aerobic detoxification and purification of wastewater streams.
It is another objective of the present invention to provide a for an aerobic bacterial process to establish a nitrogen cycle in about 1-3 days and more preferably in about 24 hours in aqueous media. Such process uses Enterobacter sakazakii and Bacillus coagulans. In another aspect these are used in combination with one or both Bacillus subtillis and Bacillus licheniformis. In another aspect, the bacteria used are limited to bacillus type bacteria.
It is a prime object of the present invention to provide for a composition and method for bacteria mediated reduction of organonitro or nitrate compounds, where the by products do not include toxic gases.
It is an important object of the present invention to provide for a bacteriochemical composition and method for bacteria reduction of organonitro or nitrate compounds, where the by-products are not malodiferous species.
It is another object of the present invention to provide for a bacterial composition and a method for aerobic oxidation of wastewater pollutants in aqueous environments such as wastewater streams, wastewater systems, aquariums, septic tanks, polluted water bodies, or for composting, including ammonia and its inorganic salts, nitrous acid and its salts, organic amines, organic and inorganic hydroxyl amines and nitroso compounds.
It is an important object of the present invention to provide for a bacterial composition that can reduce in septic tanks or other sewage treatment vessels and systems that amount of: sewage sludge, ammonia, nitrous and nitroso compounds, nitrates, phenols and phosphorous.
It is another prime objective of the present invention to provide for a composition and method for the treatment of water to reduce the ammonia, nitroso and nitrate, phosphorous content of drinking water.
It is yet another object of the present invention to provide for a composition and method for preventing eutrophication by effecting the total removal of nitrogen containing compounds, and for the total reduction of phosphorous containing compounds of aquaculture and marine systems.
It is an object of the present invention to provide for novel bacterial compositions and a method for starting and maintaining healthy, vital habitats for aqua systems, including salt water and fresh water aquariums, fish and fish hatcheries, and general marine aquacultures.
This invention is a new composition and method for the biological treatment of aqua systems, including wastewater detoxification and vital management. In particular, the present invention is directed towards a novel biological composition and method for the treatment of contaminated aqueous systems.
In one aspect the invention is use of a mixture of bacteria selected from and limited to bacillus bacteria. The process is believed to be a totally aerobic process.
In one embodiment the essential components of the present invention are the bacterial composition, and are referred to here as Mixture A. Optionally, in an additional preferred embodiment, the bacterial composition (Mixture A) is mixed with additional ingredients, which include an enzyme and living tissue solid preparations (Mixture B), and optionally a carbon platform and carbon source (Mixture D).
The bacterial composition, Mixture A, comprises bacteria from the following families: Bacillus subtillis, Bacillus sphaericus, Bacillus megatarium, Bacillus licheniformis, Enterobacter sakazakii, Bacillus coagulans, Bacillus cereus, Bacillus pasteurii, Bacillus cirroflagellosus and Bacillus pumilus. The bacterial composition of Mixture A is present in the present invention from about 45 weight percent to about 100% total composition and preferably, from about 50 weight percent to about 90 weight percent, and most preferably from about 60 weight percent to about 70 weight percent of the total composition.
The solid spores from the bacterial family of Bacillus subtillis (ATCC#465,14617,14618) are present in the composition of the present invention from about 1 weight percent to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus sphaericus (ATCC#4525,10208,12123,12300) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus megatarium (ATCC#6458,6459,8245,10778) are present in the composition of the present invention from about 1 weight percent to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus licheniformis (ATCC#6634,8480,21416) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Enterobacter sakazakii (ATCC#12868,29004) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus coagulans (ATCC#7050,15949,35670) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus cereus (ATCC#6464,9139,10702,12480) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus pasteurii (ATCC#6452645311859) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus cirroflagellosus (ATCC#14411) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The solid spores from the bacterial family of Bacillus pumilus (ATCC#7061,14884) are present in the composition of the present invention from about 1 part to about 20 weight percent, preferably about 6 weight percent to about 15 weight percent and most preferably about 10 weight percent of the total bacterial composition comprising Mixture A.
The present invention can include an enzyme composition, Mixture B, which comprises from about 0 weight percent to about 55 weight percent, preferably from about 10 weight percent to about 45 weight percent and most preferable 28 weight percent to about 40 weight percent of the mass of the total composition (Mixture A plus Mixture B) of the present invention.
Preferably the enzymes are Cellulase, Amylase, Protease and Lipase.
A suitable source of the enzyme Cellulase is present in Mixture B of the present invention up to about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent to about 90 weight percent, preferably from about 10 weight percent to about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition.
A suitable source of the enzyme Amylase is present in Mixture B of the present invention up to about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent to about 90 weight percent, preferably from about 10 weight percent to about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition.
A suitable source of the enzyme Protease is present in Mixture B of the present invention from about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent to about 90 weight percent, preferably from about 10 weight percent to about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition.
A suitable source of the enzyme Lipase is present in Mixture B of the present invention up to about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent about 90 weight percent, preferably from about 10 weight percent to about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition of the present invention referred to as Mixture B.
Also as part of Mixture B, this optionally living tissue preparation is of the pancreatic and kidney types.
A suitable source of the pancreatic preparation named Pancreatic (Hiatt Distributors Limited) is present in Mixture B of the present invention, from about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent to about 90 weight percent, preferably from about 10 weight percent to about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition of the present invention referred to as Mixture B.
A suitable source of the kidney preparation named Microbia (Hiatt Distributors Limited) is present in Mixture B of the present invention, up to about 100 weight percent of the mass of Mixture B, and also from about 1 weight percent to about 90 weight percent, preferably from about 10 weight percent to from about 35 weight percent, and most preferably from about 15 weight percent to about 25 weight percent of the mass of the composition of the present invention referred to as Mixture B.
The compositions of the present invention may be employed to treat aqueous systems. The amount of the composition (Mixture A, or Mixture A plus Mixture B) of the present invention to treat 6,000 gallons of impure water is from about 0.21 pounds to about 10.0 pounds, preferably from about 0.75 pounds to about 6.0 pounds, and most preferably from about 1.25 pounds to about 3.0 pounds.
The present invention may include a mixture C comprised of a carbon species. In the past, particulate carbon was replaced periodically when used as a filter. It has been discovered that this effectively discards much of the living bacillus which uses the carbon pellets as a biological platform. In the present invention, particulate (pellitized) carbon comprising elemental carbon is used as a food source for the bacteria ingredients. It is elemental carbon.
Also the invention uses any carbon source to the extent it is soluble in water provides a measurable TOC (total organic carbon). The portion is solution will be available as a food source for the bacteria. It is believed that the bacteria feeds on the dissolved carbon, which is further believed to continue to be produced as the cycle proceeds.
This discovery therefore calls for leaving the carbon particulates in contact with the treatment subject as a permanent bacterial platform. Thus the carbon particulates are used in liquid remediation along with the bacillus ingredients. This speeds up the denitrifying process so that completion can be accomplished in as little as one day.
Therefore, Mixture C may be incorporated in the present invention to impart nourishment to the biological aquacultures of the present invention, and to provide a growing surface for the cultivation of the bacterial colonies of the present invention. The carbon source is chosen from the group consisting of coal, coconut shell, or synthetic lignite. The amount of the carbon source is preferably about 0.1667 times the number of gallons of the treatment subject.
Mixture A is considered as being grouped into and is preferably prepared as five groups of ingredients.
Group 1 is:
Bacillus subtillis; 
Bacillus sphaericus; 
Bacillus megatarium; and
Bacillus licheniformis. 
Group 2 is:
Enterobacter sakazakii; and
Bacillus coagulans. 
Group 3 is:
Bacillus cereus. 
Group 4 is:
Bacillus pasteurii; 
Bacillus cirroflagellosus. 
Group 5 is:
Bacillus pumilus. 
The groups are prepared in the desired concentration and then mixed together.
It is normal that the bacteria will be prepared for use in a mixture with a carbon based carrier. For example bran flake (Purina) is used in concentrations of 1 part dry bacteria mixture to about 40 parts of bran flake.
For convenience, any aqueous body, wastewater stream or system or whatever body is to be treated is referred to herein as the treatment subject. Also, for convenience, the mixture of ingredients used for treatment will be referred to as the treating mixture.
It has been found that if Group 2 is omitted there is very little reduction in the ammonia content of the subject mixture compared to the use of the Group 2 ingredients.
This is taken to evidencing a synergistic affect because the ammonia oxidating organisms are in Group 1. The Group 1 ingredients are known to be active in reducing the levels of ammonia and nitrate aqueous species. The combination of Groups 1, 3, 4 and 5 show only very low levels of activity in reduction of aqueous ammonia species. The addition of Group 2 markedly accelerates not only the reduction of the amounts of aqueous ammonia species but also accelerates the rate of the consumption of all nitrogen waste in accordance with the classical nitrogen cycle.
In further particular the present method and mixture omits any nitrosomas, nitrobactor and pseudomonas or any other nitrofying non-bacillus bacteria, as was commonly used in the past, leaving a totally bacillus system having the Group 2 ingredients.
This results in faster reaction to reduce the quantity of ammonium nitrates and nitrites in a treatment subject.
The present invention therefore reduces the nitrogen cycle action from about 28 days, typically in the past, to 1-3 days. The rate of the process is temperature variable. The preferred temperature of treatment is from about 17 EC.-45 EC., most preferred being 21 EC.-40 EC. Thus this remediation treatment operates with bacillus spore cells in the absence of any vegetative cells. It is a totally aerobic process. In proper concentrations in treatment subject it can operate in 1-3 days.
Either one of Bacillus subtillis or Bacillus licheniformis can be used without the other but when both are used, an accelerated affect is seen over the use of either above such that the process can operate in as little as one day.
In one embodiment the invention is a mixture of bacillus in one particular having bacillus coagulans and absent any nitrosomas, nitrobacter, and pseudomonas providing thereby an all bacillus mixture of Bacillus subtillis, Bacillus sphaericus, Bacillus megatarium, Bacillus licheniformis, Enterobacter sakazakii, Bacillus coagulans, Bacillus cereus, Bacillus pasteurii, Bacillus cirroflagellosus and Bacillus pumilus. 
In the alternative embodiment a carbon ingredient is used in contact with the treatment subject. This can be done using a carbon canister to pass the treatment subject through, or by free addition of carbon particulates into the treatment subject. This provides both a carbon source to increase the TOC of the treatment subject and also provides a biological platform for the bacillus ingredients. The TOC functions as a food source for the bacillus.
Other inorganic toxic aqeuous bodies, wastestreams and systems have also been successfully treated with the present invention. In particular, orthophosphates, heavy metals including trivalent and hexavalent chromium, have been successfully treated.
Organic and inorganic sulphur compounds are substantially reduced in levels in a short period of time (within 24 hours).
The invention will also consume algae, both micro algae and filament algae.
The invention has been successfully used for composting.
The invention has also been successfully used for in situ bioremediation of soil.
The invention has been used for the reduction of nitrate in drinking water.
The invention can be used in salt water, urine, fresh water and brackish water. In each case the ingredients are activated in the same medium in which they will be added for treatment or are poured directly into the treatment subject.
When treating soil, the ingredients are activated in water of concentrations between about 10 to about 1,000 ppm, preferably about 50 ppm.
Water is added to the soil to be treated, and the treatment mixture is added and mixed in the soil. An accelerant may be added if desired.